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[The disulfide bridges of the trypsin-kallikrein inhibitor K from snails (Helix pomatia). Thermal inactivation and proteolysis by thermolysin (author's transl)].

Isoinhibitor K is the main component of the complex mixture of isoinhibitors of broad specificity secreted into the mucus by the Roman snail (Helix pomatia). The disulfide pairing was determined after the amino acid sequence had been elucidated. Two cystine-containing peptides with the disulfide bridges Cys32-Cys53 and Cys32-Cys53 plus Cys7-Cys57 were obtained after thermolytic hydrolysis of the native inhibitor at 80 degrees C and chromatographic separation of the peptides using SE-Sephadex. The Cys16-Cys40 disulfide bridge could be reduced selectively by sodium borohydride with no loss in biological activity. This property and the covalent structure correspond to that of the intracellular inhibitor from bovine organs, which is largely homologous in its amino acid sequence to the secretory inhibitor from the snail. The complete covalent structure of isoinhibitor K will be presented. The snail inhibitor is less stable against proteolytic inactivation by thermolysin and against thermal denaturation at pH 8.0 than the inhibitor from bovine organs (Kunitz inhibitor).

Amino Acid Sequence

[Long-term results of biological control by predation. Experimental study of population development of several species of snails (author's transl)].

The evolution of malacological fauna, which is subject to prediation of Zonitidae snails, is studied on two types of habitats of Lymnaea truncatula: swampy meadows and banks of river maintained by a periodical grass-mowing. Two years are necessary for obtaining an entire elimination of Lymnaea truncatula. Then other species of Lymanaeidae and Succinea are affected by predation, but Succinea snails do not pass away completely. The other snails are not affected or little by predation.

Animals

The role of Chaetogaster limnaei in the dynamics of trematode transmission in natural populations of freshwater snails.

Chaetogaster limnaei, an oligochaete was always found infesting Bulinus (Phy.) globosus, B.(B.) forskalii and Lymnaea natalensis in Bo, Sierra Leone, living below the shell or embedded in the mucus of the foot. The worms had no preference for snails with trematode infection. The population was highest in the dry season when fewer snails were infected by trematodes. This is suggested to be a result of the protection the worms give to snails against invading miracidia.

Animals

Enzyme analyses of Bulinus africanus group snails (Mollusca: Planorbidae) from Tanzania.

38 population samples of snails of the Bulinus africanus group, collected from three separate areas of Tanzania, have been examined. Enzymes in crude digestive gland extracts of individual snails have been analysed by isoelectric focusing in polyacrylamide gels. The enzymes studied were: malate dehydrogenase (MDH); phosphoglucomutase (PGM); glucosephosphate isomerase (GPI); acid phosphatase (AcP) and hydroxybutyrate dehydrogenase (HBDH). Samples of B. nasutus were clearly differentiated from other species and enzyme differences were apparent between samples from the lake and coastal areas. Similarly, although clear distinctions could not always be made, samples of B. africanus, B. globosus and B. ugandae were characterized by their enzyme types. Individual variation was detected within populations and the significance of enzyme polymorphisms in relation to identification has been considered. No correlation was found between snail enzyme type and susceptibility to Schistosoma haematobium or S. bovis.

Acid Phosphatase

A demonstration of the presence of anti-snail antibodies in individuals infected with Schistosoma haematobium.

An extract of the snail Bulinus (Physopsis) africanus, the intermediate host of Schistosoma haematobium, was used as the antigen in haemagglutination tests in a survey of the prevalence of anti-snail antibodies in various population groups. It was found that sera from known bilharzia-infected individuals and randomly selected individuals from bilharzia endemic areas had significantly higher incidences as well as higher titres of antibodies to this snail antigen than non-infected individuals and individuals from non-endemic areas.

Adult

Interference with Fasciola hepatica snail finding by various aquatic organisms.

Previous studies using radioactive miracidia have shown that a number of non-host snails and bivalves, interposed as 'decoys' in linear test channels, may interfere with the capacity of Fasciola hepatica miracidia to infect Lymnaea truncatula. Applying similar experimental principles, the role of several other potential interferents have been analysed in the present study. Daphnia pulex (Cladocera) and larvae of Corethra sp. (Diptera) exercised significant interfering effects by protecting 'target' snails from infection. Evidence suggested that this effect was a result of their normal predatory behaviour. Other organisms including Herpobdella testacea and Helobdella stagnalis (Hirudinea), Acellus aquaticus (Isopoda), Planaria lugubris (Turbellaria) and L. truncatula egg clusters failed to interfere with miracidial host-finding. Nor did P. lugubris and L. truncatula 'conditioned water' interfere with the capacity of the miracidia to infect their host snail.

Animals

The influence of stream geology on the distribution of the bilharzia host snails, Biomphalaria pfeifferi and Bulinus (physopsis) sp.

The Gladdespruit and Komati River surveys have shown that similar environmental conditions have developed where the watercourses flowed over particular rock types. In these watercourses a potentially useful association was found between the occurrence of permanent, lentic habitats produced by the weathering of bedrock with a hardness above 5 in Mohs' Scale of Hardness and the longitudinal distribution of persistent populations of the bilharzia intermediate host snails Biomphalaria pfeifferi and Bulinus (Physopsis) sp. Bilharzia transmission follows this pattern in these waterways. Weathering of bedrock with hardness below 5 in Mohs' scale, and/or flow over beds of alluvial sand, produced perpetually lotic (flowing) environments without persistent host snail populations because of the snail's inability to tolerate high current speeds. However a few temporary populations survived in temporarily lentic habitats during the dry winter season.

Africa

Schistosoma mansoni: identification of chemicals that attract or trap its snail vector, Biomphalaria glabrata.

A new bioassay for chemical attractants of aquatic snails demonstrated that Biomphalaria glabrata could be attracted to or trapped in the vicinity of homogenates of lettuce. Fractionation of homogenates revealed the amino acids glutamate and proline and the primary attractants. Attraction was specific for the L form of glutamate. Proline appeared to stimulate reproductive activity. Glutathione, gamma-aminobutyric acid, and a number of other compounds had no effect. Extracts of lyophilized snail tissue also attracted other snails and may thus contain pheromones. These results permit formulation and testing of controlled-release attractants designed to overcome the repellant effects of slow-release molluscicides, as well as the design of stimulants to be used with no-release poisons.

Amino Acids

The chemical ecology of Biomphalaria glabrata: the effects of ammonia on the growth rate of juvenile snails.

When juvenile specimens of Biomphalaria glabrata were subjected to concentrations of ammonia ranging from 1-100 mug/ml in various media the following effects were observed: the addition of ammonia to borate buffered media caused mortality. Both borate and tris-buffered media caused a decrease in the growth rate of snails when compared with controls in SSW. The growth rates of the snails could be enhanced by increasing the concentration of ammonia to critical thresholds, but further increases beyond these thresholds resulted in growth inhibition. The toxicity of ammonia in ambient water was augmented by an an increase in pH. The possible causation and ecological significance of these effects are discussed. There are indications that the snails are physiologically well-adapted to utilize ammonia when required and also to control its excretion and uptake from the medium.

Ammonia

Seasonal population densities of snails transmitting urinary and intestinal schistosomiasis in Lusaka, Zambia.

The two molluscan intermediate hosts of S. haematobium schistosomiasis: Bulinus africanus and Bulinus globosus, were found most commonly in habitats that retained water for a substantial part of the year rather than in temporary rain-filled pools. Biomphalaria pfeifferi which transmits S. mansoni were found principally in streams and impoundments like dams but scarce in rivers. In a permanent habitat, the peak density of snails was between the warm, dry months of August and September while in a temporary habitat B. (Physopsis) sp. were most abundant towards the end of the rainy season in March and April. The influence of rainfall and the prevailing the temperature on the fluctuations in the density of snails populations is considered. Further, the low infection rate in snails found in the field is discussed in relation to the critical effect of temperature on this feature. The problem of increasing migration of rural inhabitants to the peri-urban areas and the possible escalation of prevalence rates is considered in relation to the control measures that may be instituted in view of the substantial body of base-line data now available to effect focal control.

Animals

Failure of irradiated Echinostoma audyi and Hypoderaeum dingeri to sensitize Lymnaea rubiginosa snails.

Attempts to induce acquired resistance in Lymnaea rubiginosa snails against the echinostomes Echinostoma audyi and Hypoderaeum dingeri by means of irradiated miracidia were unsuccessful, although Lie and coworkers using similar methods had recently sensitized Biomphalaria glabrata against Echinostoma lindoense (1975a). In contrast to the B. glabrata-E. lindoense system, in Lymnaea rubiginosa the amebocytic response to irradiated parasites was slow: irradiated E. audyi sporocysts were encapsulated 15 to 28 days and H. dingeri sporocysts 20 to 27 days postexposure. No obvious enlargement of the amebocyte-producing organ was seen. No resistance was demonstrable to subsequent homologous challenge. Development of acquired resistance to a trematode infection in snails may be related to the speed with which the snails destroy the irradiated sporocysts.

Animals

Ultrastructural study of the shell-repair membrane in the snail, Helix pomatia L..

The shell-repair membrane of the snail, Helix pomatia, has been studied with the transmission electron microscope (TEM). The ultrastructure of the repair membrane, in the initial stages of calcification, revealed the presence of a fibrillar protein, proteoglycan granules, osmiophilic vesicles, and cytoplasmic dense bodies of different size and structure. The involvement of the cell constituents in the formation of calcifying centra and initial crystal formation is discussed. The amoebocytes present within the repair membrane appeared to be involved in three different functions: (1) phagocytosis, (2) release of granules, vesicles and dense bodies, and (3) secretin of a fibrillar protein. The possible lytic function of the amoebocytes is mentioned. The common features in the minearlizing process of the shell-repair membrane of the snail and the epiphyseal cartilage of the mammals were noted.

Animals

The amino-acid sequence of isoinhibitor K form snails (Helix pomatia). A sequence determination by automated Edman degradation and mass-spectral identification of the phenylthiohydantoins.

Performic-acid-oxidized isoinhibitor K of snails (Helix pomatia) was subjected to arginine-directed tryptic proteolysis. Six peptide fragments including one overlap peptide from limited cleavage of the Arg-3-Pro-4 bond were purified to homogeneity. Four arginine peptides and the C-terminal peptide were sequenced by automatic Edman degradation using a special peptide program. The phenylthiohydantoins were all identified by chemical ionization mass spectrometry, except four cysteic acid residues that were identified on an amino acid analyzer after acid hydrolysis. Quantitative evaluation of the phenylthiohydantoins by chemical ionization mass spectrometry using total molecular-ion beam integration greatly facilitated sequencing. The mass spectrum of the dipeptide less than Glu-Gly revealed that the N-terminus was blocked by pyroglutamic acid. The complete amino acid sequence of isoinhibitor K was determined. An almost 50% homology between the sequences of the snail inhibitor and the bovine trypsin-kallikrein inhibitor (Kunitz) became obvious. A comparison of all homologous sequences of this particular class of proteins known to date is presented.

Amino Acid Sequence

The moroccan food snail, Helix aspersa, as a source of Salmonella.

A total of 270 samples, nine lots of 30 samples each, of imported Moroccan food snails was examined for the presence of Salmonella. Eighty-four samples (an overall incidence of 31.11%) and all nine lots contained Salmonella. No significant difference (P greater than 0.25) in the number of positive samples was observed by using either selenite cystine both or tetrathionate broth when the samples had been pre-enriched in lactose broth. When used as direct selective enrichments with samples not pre-enriched in lactose broth, tetrathionate broth was significantly (P less than 0.05) more productive than selenite cystine broth. The overall detection of Salmonella-positive samples by direct enrichment was significantly greater (P less than 0.001) than by pre-enrichment. A variety of uncommon serotypes occurrence and incidence, and the concomitant human health potential, of Salmonella in one species of live, imported food snails.

Culture Media

Enzyme changes accompanying thermal acclimation in two species of pleurocerid snails.

Goniobasis cahawbensis is a stream snail that experiences an annual temperature cycle. G. cochliaris is limited in distribution to springs, and their immediate vicinities, which are characterized by nearly constant annual temperatures. The present study sought to determine whether temperature dependent biochemical differences exist that might account for the differential distribution of these congeneric pleurocerid snails. Eight enzymes were examined following acclimation to 10 degrees, 17 degrees and 24 degrees C. No significant temperature dependent qualitative differences in enzyme phenotypes were demonstrable in either species by starch-gel electrophoresis for malate dehydrogenase, glucose-6-phosphate dehydrogenase, phosphogluconate dehydrogenase, phosphoglucomutase, superoxide dismutase and acetyl and butyryl esterases. Significant quantitative differences were observed in three of these enzymes. G. cahawbensis cytosol malate dehydrogenase activity increased significantly with increasing acclimation temperature, while G. cochliaris malate dehydrogenase activity remained unchanged. The activities of glucose-6-phosphate dehydrogenase did not differ significantly between acclimation temperatures for either species; however, the overall activity of both enzymes was significantly higher for G. cochliaris. Appreciable levels of LDH activity were not demonstrable by electrophoresis or enzymatic assay.

Adaptation, Physiological

Water regulation in aestivating snails. Ultrastructural and analytical evidence for an unusual cellular phenomenon.

Aestivating snails form abundant lamellate vesicles in the cells of the mantle collar, an epithelium known to regulate the rate at which water is lost from its surface. Since lamellate vesicles are much reduced in hydrated mantle tissue of recently stimulated animals it is tentatively concluded that the vesicles, and their contents, form a barrier to water movement within these cells. X-ray microanalysis of unfixed thin sections shows that there is a concentration gradient of ions within these cells in aestivating animals which is not present in stimulated snails.

Adaptation, Physiological

The penetration of Fasciola hepatica miracidia into the snail host Fossaria bulimoides. A scanning electron microscope study.

Snails of the species Fossaria bullimoides were exposed to miracidia of Fasciola hepatica for given periods of time. The course of penetration was followed by means of scanning electron microscopy. Topographical features of miracidial morphology were studied with emphasis on epidermal plates and body shape. Comparisons are made on the rate of penetration into this snail host.

Animals